A novel human type II cytokeratin, K6hf, specifically expressed in the companion layer of the hair follicle

In an attempt to identify new members of the human type II hair keratin family by means of 3'- and 5'-RACE methods and cDNA from anagen hair follicles, we detected a sequence that encoded a hitherto unknown type II cytokeratin. The novel cytokeratin comprises 251 amino acids and exhibits the highest sequence homology with K5. Comparative one- and two-dimensional western blots of keratins from anagen hair bulbs, containing or not containing the outer and inner root sheaths (ORS/IRS), and from footsole epidermis with an antibody against the new cytokeratin, revealed its comigration with K6 and its expression in the ORS/IRS complex. We have therefore named the new cytokeratin K6hf, to distinguish it from the various K6 isoforms and to indicate its expression in the hair follicle. Both in situ hybridization with a K6hf-specific cRNA probe and indirect immunofluorescence with the K6hf antibody showed that K6hf is exclusively expressed in the so-called "companion layer" of the hair follicle, a single layered band of flat and vertically oriented cells between the cuboidal ORS cells and the IRS that stretches from the lowermost bulb region to the isthmus of the follicle. Concomitant K17 and K16 expression studies showed that besides suprabasal ORS cells, these cytokeratins are sequentially expressed subsequent to K6hf in companion cells above the hair bulb. Our study confirms the view of a vertically oriented companion layer differentiation. The clearly delayed K17 and K16 expression relative to that of K6hf in companion cells most probably excludes these keratins as possible type I partners of K6hf and suggests the existence of a still unknown type I partner of its own. Thus, not only morphologically but also biochemically, the companion layer is different from the ORS and can therefore be regarded as an independent histologic compartment of the hair follicle.     

Proline isomerization-independent accumulation of an early intermediate and heterogeneity of the folding pathways of a mixed alpha/beta protein, Escherichia coli thioredoxin

Oxidized Escherichia coli thioredoxin (Trx) is a small protein of 108 residues with one disulfide bond (C32-C35 essentially involved in the activity) and no prosthetic moieties, which folds into a structural motif containing a central twisted beta-sheet flanked by helices that is found in many larger proteins. The kinetics of refolding of Trx in vitro have been investigated using a newly developed active site titration assay and continuous or stopped-flow (SF) methods in conjunction with circular dichroism (CD) and fluorescence (Fl) spectroscopy. These studies revealed the presence of early folding intermediates with "molten globule or pre-molten globule" characteristics. Measurements of the ellipticity at 222 nm indicated that about 68% of the total change associated with refolding occurred during the dead time (4 ms) of the stopped-flow instrument, suggesting the formation of substantial secondary structure. The reconstruction of the far-UV CD spectrum of the burst intermediate using combined continuous and stopped-flow methods showed the formation of a defined secondary structure that contains more beta-structure than the native state. Kinetic measurements using SF far-UV CD and Fl over a wide range (0.087-6 M) of GuHCl concentrations at two temperatures (6 and 20 degreesC) demonstrated that the population formed during the 4 ms dead time contained multiple species that are stabilized mainly by hydrophobic interactions and undergo further folding along alternative pathways. One of these species leads directly and rapidly to the native state as demonstrated by active site titration, while the two others fold into a fourth intermediate that is slowly converted to the native protein. Double-jump experiments suggest that the heterogeneity in folding behavior results from proline isomerizations occurring in the unfolded state. Conversely, the accumulation of the burst intermediate does not depend on proline isomerizations.     

Structural Induction and Coinduction in a Fibrational Setting

We present a categorical logic formulation of induction and coinduction principles for reasoning about inductively and coinductively defined types. Our main results provide sufficient criteria for the validity of such principles: in the presence of comprehension, the induction principle for initial algebras is admissible, and dually, in the presence of quotient types, the coinduction principle for terminal coalgebras is admissible. After giving an alternative formulation of induction in terms of binary relations, we combine both principles and obtain a mixed induction/coinduction principle which allows us to reason about minimal solutionsX≅σ(X) whereXmay occur both positively and negatively in the type constructor σ. We further strengthen these logical principles to deal with contexts and prove that such strengthening is valid when the (abstract) logic we consider is contextually/functionally complete. All the main results follow from a basic result about adjunctions between “categories of algebras” (inserters).     

Developments in the treatment of alcoholism

The treatment of alcoholism has changed during the past 2 decades. Notable developments have occurred in pharmacotherapy, psychotherapy, and health-care delivery. A better understanding of the biologic basis for addiction has led to clinical trials of medications that target neuroreceptors. One such medication is the opiate antagonist naltrexone, which decreases the craving for alcohol. Psychosocial interventions continue to be the mainstay of alcohol treatment programs. The efficacy of three different therapies was demonstrated in a study called Project MATCH (Matching Alcoholism Treatments to Client Heterogeneity). This study, however, did not prove the patient-treatment "matching" hypothesis. In addition to therapies provided by addiction specialists, interest is growing in the use of brief motivational techniques in primary-care settings. As the field of addiction responds to an unfolding health-care delivery system, a broader range of treatment options in conjunction with a greater opportunity to individualize patient care is evolving.     

Bound-abrasive polishers for optical glass

Polishing abrasives that have been bound in a solid matrix can offer several potential advantages over loose-abrasive processes with pitch or polyurethane laps for finishing of optics. These advantages include polishing efficiency, temperature stability, cost of consumables, and compatibility with computer numerically controlled generating machines. Unfortunately, little has been published on bound-abrasive polishers, and very few commercially available products exist. We have developed several compositions and manufacturing techniques that show promise for polishing a variety of optical glasses. We establish the various criteria for a successful bound-abrasive polisher. The numerous variables to be considered in designing these polishers are identified, and the results of screening experiments are used to find successful compositions. Polishing experiments with bound abrasives in arrays of pellets, as ring tools, and as full-contact laps are described.     

Homogeneous asymmetric hydrogenation ofo-chloroacetophenone to enantiomerically pureo-chloro-(1-phenyl)ethanol catalyzed by BINAP-Ru(II) complex

The BINAP-Ru(II) catalyst (2,2-bis(diphenylphosphino)-1,1-binaphthyl)chloro(p-cymene)rutheniun chloride is found to be highly active and enantioselective for homogeneous asymmetric hydrogenation of o-chloroacetophenone to optically pureo-chloro-(1-phenyl)ethanol. In contrast to results of Noyori and coworkers, no organic and inorganic bases are required to obtain high activities and enantioselectivities     

Surface microroughness of optical glasses under deterministic microgrinding

Deterministic microgrinding of precision optical components with rigid, computer-controlled machining centers and high-speed tool spindles is now possible on a commercial scale. Platforms such as the Opticam systems at the Center for Optics Manufacturing produce convex and concave spherical surfaces with radii from 5 mm to ∞, i.e., planar, and work diameters from 10 to 150 mm. Aspherical surfaces are also being manufactured. The resulting specular surfaces have a typical rms microroughness of 20 nm, 1 μm of subsurface damage, and a figure error of less than 1 wave peak to valley. Surface roughness under deterministic microgrinding conditions (fixed infeed rate) with bound abrasive diamond ring tools with various degrees of bond hardness is correlated to a material length scale, identified as a ductility index, involving the hardness and fracture toughness of glasses. This result is in contrast to loose abrasive grinding (fixed nominal pressure), in which surface microroughness is determined by the elastic stiffness and the hardness of the glass. We summarize measurements of fracture toughness and microhardness by microindentation for crown and flint optical glasses, and fused silica. The microindentation fracture toughness in nondensifying optical glasses is in good agreement with bulk fracture toughness measurement methods.     

Flap prefabrication: effectiveness of different vascular carriers

A new experimental model of a vascular carrier to prefabricate a "secondary" island flap, the popliteal musculovascular pedicle, was developed in the rat. Using quantitative skin-surface fluorometry 30 minutes after sodium fluorescein injection and a flap survival area in the prefabricated 8 x 2.5-cm abdominal composite island flap, we compared the revascularization ability of our muscular carrier to nonrevascularized controls: the skeletonized arteriovenous pedicle and the fasciovascular pedicle. The free composite graft with no vascular carrier exhibited near-total necrosis. The skeletonized vascular pedicle demonstrated 15.2% +/- 7.8% perfusion of normal skin on dye fluorescence index measurements and 50% flap survival. The fasciovascular pedicle exhibited better revascularization, with a dye fluorescence index of 36.2 +/- 15.5 (p < 0.01) and 90% +/- 10% flap survival (p < 0.001). India ink injection study and histological examination of our model provided visual evidence of revascularization from the musculovascular pedicle, along with preservation of the carrier's muscular architecture. The musculovascular pedicle is a reliable carrier for making new, vascularized composite flaps.     

The gradual threshold model of ambivalence: relating the positive and negative bases of attitudes to subjective ambivalence

Eighty subjects viewed and visually imagined upright or rotated alphanumeric characters and later judged whether test characters were previously seen or imagined (reality monitoring). Identification and test characters were presented verbally or visually. When characters were identified and tested verbally, source confusions (misjudging a seen character as "imagined" and vice-versa) were infrequent and were comparable for rotated and upright characters. When characters were identified and tested visually, source confusions were more frequent and were influenced by character rotation. Memories for imagined characters were especially susceptible to source confusion. Also source confusions for seen characters increased when characters were rotated. These results are consistent with the proposal that increasing sensory similarity between perceived and imagined items increases source confusion and that perceived rotation generates cognitive operations similar to those generated when the subject imagines a character rotated.     

Automatic verification of Java programs with dynamic frames

Framing in the presence of data abstraction is a challenging and important problem in the verification of object-oriented programs Leavens et al. (Formal Aspects Comput (FACS) 19:159–189, 2007). The dynamic frames approach is a promising solution to this problem. However, the approach is formalized in the context of an idealized logical framework. In particular, it is not clear the solution is suitable for use within a program verifier for a Java-like language based on verification condition generation and automated, first-order theorem proving. In this paper, we demonstrate that the dynamic frames approach can be integrated into an automatic verifier based on verification condition generation and automated theorem proving. The approach has been proven sound and has been implemented in a verifier prototype. The prototype has been used to prove correctness of several programming patterns considered challenging in related work.